Assay analysis

ABSTRACT

Apparatus and assemblies for the detection of at least one analyte in a sample are shown and described. In one embodiment, the assembly generates a test result from an assay and includes an integrated reader and incubator, wherein the incubator incubates the assay as the reader generates the test result. The reader typically has an optical detector aligned with a light source for detecting a plurality of transmission of light on the assay. The result is systems and methods to improve the detection of the presence and/or absence of at least one analyte in a sample.

This application is a Continuation of U.S. Pat. No. 10,254,233 filedApr. 20, 2018, and is based on and claims priority to U.S. applicationSer. No. 13/819,064 filed Nov. 11, 2013; PCT/US2011/49170 filed Aug. 25,2011; U.S. Provisional Application No. 61/454,771 filed Mar. 21, 2011;and U.S. Provisional Application 61/377,287 filed Aug. 26, 2010, all ofwhich are incorporated herein by reference in their entireties.

FIELD OF THE TECHNOLOGY

The present disclosure relates generally to analytical testing, and moreparticularly to improved test strips for the detection of an analyte.

BACKGROUND

Reagent strips and films are often a helpful analytical tool in thefields of clinical chemistry, analytical medicine, and food sanitationdiagnostics. For example, it is advantageous to determine or to test,through quantitative or qualitative methods, various matrices, includingbody fluids such as serum and urine, and food, such as meat products,fruit, vegetables, milk, honey and the like. Such matrices can be testedfor a variety of analytes including a variety of chemicals, biochemicalsand biological molecules such as bacteria, antibiotics, for example,sulfa drugs, tetracyclines, beta-lactam drugs; toxins, such asaflatoxin, zearalonone, ochratoxin, T-2, and vomitoxin, pesticides suchas organophosphates and carbamates, and active metabolites, either inmaterials or on the surface of materials or a combination thereof.

Generally, lateral flow assays are membrane-based test devices in whicha sample that is suspected of containing the analyte of interest isplaced at or near one end of the membrane strip. The sample is carriedto the opposite end of the membrane strip by mobile phase that traversesthe membrane strip, for example by capillary action. While traversingthe membrane strip, the analyte in the test sample, if any, encountersone or more reagents. The reagents can include binders for the analyte.Binders can be mobile and, therefore, flow with the sample, or beimmobilized on the test strip as a capture agent. Depending on the testconfiguration, either the analyte binder, the analyte itself, or someother reagent in the test system will be captured by the immobilizedcapture agent and, thereby, produce a detectable signal. The signal canbe generated by a label provided within the assay. The detectable signalcan be measured, such as by an optical reader.

The presence and, in some cases, the concentration, of an analyte on areagent strip may be determined by measuring the optical reflectancefrom an area of development on the strip. For example, the area ofdevelopment on the strip may be an area of color development. Percentreflectance can be used to determine the result.

Testing commonly occurs in a controlled environment, such as alaboratory, but testing in non-laboratory settings is also common. Insome applications speed and ease of use is particularly important. Forexample, in food processing it would be advantageous for tests to be runin non-laboratory settings because processors must wait for results.Further, it would also be advantageous for tests to be run on trucksduring transport of the items. For that reason, it would be advantageousto accelerate the speed of testing, reduce the cost of equipment andtests, improve the ruggedness of the apparatus, and enhance the ease ofuse and simplicity of operation. In addition, it is advantageous to haveconfidence that test results are valid. Therefore, systems, methods anddevices herein also assist in preventing fraudulent use of pre-run,known negative assays in place of true samples or use of assayspre-marked to provide a negative result that does not reflect the truenature of the sample. It is also desirable to increase the ruggedness ofthe assays, systems and test procedures.

Therefore, Applicants desire systems and methods for analyte detectionwithout the drawbacks presented by traditional lateral flow assaysystems and methods.

SUMMARY

This disclosure provides improved test strips for analyte detection. Inone embodiment, a lateral flow assay for the detection of an analyteincludes a surface reflectance profile that is generally adapted toenable monitoring the test strip prior to the detection of the analyte.The reflectance profile may include a reference area to monitor apre-flow development and a test result reference area to monitor apre-test detection. Typically, at least one flow reference area isadapted to monitor a pre-flow development along the assay. Further, atleast one test result reference area is typically adapted to monitor apre-test detection of the analyte on the assay.

In particular examples, the reflectance profile includes a theoreticallight reflectance measurement. The theoretical light reflectancemeasurement may include a no-flow development theoretical value. In someexamples, the no-flow development value is a reflectance value of about85. For instance, a reflectance value of greater than about 85 generatesa signal to deactivate the detection of the analyte.

In other examples, the flow reference area includes at least onedownstream flow reference line. The downstream flow reference line mayinclude a theoretical reflectance value after the flow reference linereceives reagent flow thereon. Further, the flow reference area mayinclude an intermediary flow reference line and a downstream flowreference line. In some examples, the intermediary flow reference linemay include a theoretical reflectance value after the flow referenceline receives reagent flow thereon. The theoretical light reflectancemeasurement may include a no-analyte pre-test development theoreticalvalue. The test result reference area may include at least one test linehaving a theoretical reflectance value. Further, the test resultreference area includes at least one control line having a theoreticalreflectance value.

In yet additional examples, the test result reference area includes atleast one test line having a theoretical reflectance value and at leastone control line having a theoretical reflectance value. Typically, apre-set difference between the at least one test line's theoreticalreflectance value and the at least one control line's theoreticalreflectance value activates a test result. Further, a pre-set differencebetween the at least one test line's theoretical reflectance value andthe at least one control line's theoretical reflectance value typicallytriggers an error. For instance, the error may withhold a test result.

In other embodiments, a lateral, capillary-flow elongated test strip mayinclude at least one reagent for the detection of at least one analytein a sample. The capillary-flow elongated test strip may include a testzone, a control zone and a surface having a reflectance profile.Typically, the test zone includes immobilized thereon a test zonecapture agent that is generally adapted for capturing the at least onereagent. Further, the control zone typically includes at least onecontrol zone capture agent having a different binding affinity for theat least one reagent. The surface may have a reflectance profile that isgenerally adapted to monitor the test strip continuously, for instanceuntil the analyte is detected. Typically, the test strip generates adetectable signal upon detection of the analyte in the sample.

In some examples, the test strip may include a coding system having atleast one reference code with a corresponding testing sequence. Thetesting sequence may include at least one testing parameter, forinstance a temperature adjustment parameter, an optical reader testparameter, a reader channel selection, a combination thereof and thelike. In particular examples, the reader test parameter includes anassociated feature chosen from a standard curve, a does-response curveand a combination thereof.

In certain examples, the coding system includes a color matrices that isgenerally associated with a corresponding diagnostic test. The codingsystem may include a code chosen from a bar code, an RFID tag, acombination thereof and the like. In yet other examples, the test stripincludes a peel strip to introduce sample onto the sample absorbingmaterial, and wherein the peel strip includes a color peel tab at oneend of the peel strip associated with the color matrices of thecorresponding diagnostic test. The test strip may include an opposedsecond end having a reactor detector material. Typically, the test stripis adapted for selecting the detection of a diagnostic test group chosenfrom an antibiotic analyte, toxic analyte, analyte class and combinationthereof. The test strip may be sized and adapted to be enclosed within atest strip cavity. For instance, the test strip may be sized and adaptedto be enclosed within a test strip cavity of a removable incubationmodule.

In some examples, the test zone includes at least one analyte referenceline having a theoretical reflectance value. The theoretical reflectancevalue may be associated with a flow parameter on the test strip. Inaddition, the test zone may include a first analyte reference linehaving a first theoretical reflectance value and a second analytereference line having a second theoretical reflectance value. Thecontrol zone may include at least one control line having a theoreticalreflectance value. The theoretical reflectance value may be an opticalreflectance value. The control zone may include a first control linehaving a first theoretical reflectance value and a second control linehaving a second theoretical reflectance value.

In yet an alternative embodiment, a lateral, capillary-flow elongatedtest strip includes a test zone, control zone, a surface having areflectance profile and a coding system. Typically, the test zoneincludes immobilized thereon a test zone capture agent adapted forcapturing at least one binder. Further, the control zone typicallyincludes at least one control zone capture agent, for instance at leastone control zone capture agent having a different binding affinity forthe at least one binder. The coding system includes at least one codingsignal to correspond to a testing sequence to characterize the teststrip.

In another embodiment, an assay analysis device may include at least onetest line and at least one control line, and whereby the theoreticalreflectance value is a comparison between a reflectance value at thetest line and a reflectance value at the control line. A reflectancevalue on the assay that is inconsistent with the theoretical reflectancevalue may indicate an inadequate flow on the assay. The inadequate flowmay trigger a detectable signal to generate a no-result response. Thereflectance value on the assay that is inconsistent with the theoreticalreflectance value may indicate a prior analyte development on the assay.The reflectance values may suggest prior analyte development may triggera detectable signal to deactivate the assay measurement apparatus. Thereflectance value on the assay that is inconsistent with the theoreticalreflectance value may indicate a contaminated optical path.

A reference coding may activate a corresponding diagnostic test in theoptical detector. A multichannel reader and the reference coding mayactivate a corresponding channel in the multichannel reader. Theapparatus may include an incubator and the reference coding may activatea corresponding incubation temperature. An instruction for generating atest result may correspond to an image detection on the assay. The imagedetection may be an optical reflectance value. The assay may include atleast one test line and at least one control line, and whereby theoptical reflectance value is a comparison between a reflectance value atthe test line and a reflectance value at the control line. The apparatusmay be adapted to perform a continuous image detection of the assay. Theassay may be a lateral flow assay. For instance, the assay may be alateral, capillary-flow, elongated test strip. Further, the apparatusmay include a means for a power source.

In yet another embodiment, a lateral flow assay for the detection of ananalyte and having a test zone and a control zone, a surface having areflectance profile includes at least one flow reference and at leastone test result reference. The at least one flow reference area may beadapted to enable monitoring of a pre-flow development along the assay.The at least one test result reference area may be adapted to enablemonitoring a pre-test detection of the analyte on the assay.

The reflectance profile may include a theoretical light reflectancemeasurement. The theoretical light reflectance measurement may comprisea no-flow development theoretical value. The no-flow development valuemay be a reflectance value of about 85. A reflectance value of greaterthan about 85 may generate a signal to deactivate the detection of theanalyte. The flow reference area may include at least one downstreamflow reference line. The downstream flow reference line may include atheoretical reflectance value after the flow reference line receivesreagent flow thereon. The flow reference area may include both anintermediary flow reference line and a downstream flow reference line.The intermediary flow reference line may include a theoreticalreflectance value after the flow reference line receives reagent flowthereon. The theoretical light reflectance measurement may comprise ano-analyte pre-test development theoretical value. The flow referencemay also be the control zone.

The test result reference area may include at least one test line havinga theoretical reflectance value. The test result reference area mayinclude at least one control line having a theoretical reflectancevalue. The test result reference area may include at least one test linehaving a theoretical reflectance value and at least one control linehaving a theoretical reflectance value. A pre-set difference between theat least one test line's theoretical reflectance value and the at leastone control line's theoretical reflectance value may activate a testresult. Further, a pre-set difference between the at least one testline's theoretical reflectance value and the at least one control line'stheoretical reflectance value may trigger an error. The error maywithhold a test result.

In other embodiments, a lateral, capillary-flow elongated test stripincludes a test zone, a control zone and a surface having a reflectanceprofile. The lateral, capillary-flow elongated test strip may have atleast one reagent for the detection of at least one analyte in a sample.The test zone may include immobilized thereon a test zone capture agentthat is adapted for capturing the at least one reagent. The control zonemay include at least one control zone capture agent having a differentbinding affinity for the at least one reagent. The reflectance profilemay be adapted to enable monitoring of the test strip continuously untilthe detection of the analyte. Typically, the test strip generates adetectable signal for detecting the analyte in the sample. In someexamples, inadequate control line development, for instance according toreflectance and/or transmission at the control line, may trigger anerror. In these examples, the error may trigger a signal to generate ano-result response.

The test strip may comprise a coding system having at least onereference code with a corresponding testing sequence. The testingsequence may include at least one temperature adjustment parameter.Further, the testing sequence may include an optical reader testparameter. The optical reader test parameter may include a readerchannel selection. The reader test parameter may include an associatedfeature chosen from a standard curve, a does-response curve and acombination thereof. The reader test parameter may include at least oneassociated positive control point and at least one associated negativecontrol point. The coding system may include a color matrices. The colormatrices may include a color chosen from red, blue, green andcombination thereof. The color matrices may be associated with acorresponding diagnostic test. The coding system may include a bar code.The coding system may include an RFID tag.

The test strip may include a first end having a sample absorbingmaterial. The test strip may include a peel strip to introduce sampleonto the sample absorbing material. The peel strip may include a peeltab at one end of the peel strip to facilitate movement of the peelstrip. The sample absorbing material may be adapted to receive about 0.1to about 1.0 mL of a fluid. The sample absorbing material may comprise adry cellulosic material. Further, the test strip may include an opposedsecond end having a reactor detector material. The test strip mayinclude a releasing area having a mobile phase receptor for the at leastone analyte. The test strip may be sized and adapted to be enclosedwithin a test strip cavity. Further, the test strip may be sized andadapted to be enclosed within a test strip cavity of a removableincubation module. Typically, the test strip is adapted for selectingthe detection of a diagnostic test group chosen from an antibioticanalyte, toxic analyte, analyte class, a combination thereof and thelike.

The test zone may include at least one analyte reference line having atheoretical reflectance value. The theoretical reflectance value may beassociated with a flow parameter on the test strip. The test zonesurface may include a first analyte reference line having a firsttheoretical reflectance value and a second analyte reference line havinga second theoretical reflectance value. The control zone surface mayinclude at least one control line having a theoretical reflectancevalue. For instance, the theoretical reflectance value may be an opticalreflectance value. The control zone may include a first control linehaving a first theoretical reflectance value and a second control linehaving a second theoretical reflectance value. In some examples, thereflectance profile is adapted to enable monitoring of the test stripprior to the detection of the analyte. Further, the test result may bedetected within about thirty to about sixty seconds.

In yet another embodiment, a lateral, capillary-flow elongated teststrip includes a test zone including immobilized thereon a test zonecapture agent adapted for capturing at least one binder, a control zoneincluding at least one control zone capture agent having a differentbinding affinity for the at least one binder, a surface having areflectance profile adapted to enable monitoring of the test strip and acoding system having at least one coding signal, for instance a codingto correspond to a testing sequence to characterize the test strip. Thereflectance profile may include at least one flow reference area adaptedto enable monitoring of a flow development along the assay, and at leastone monitor reference area adapted to enable monitoring of detection ofthe analyte on the assay. In some examples, the test strip may include afirst end having a sample absorbing material. The test strip may includea peel strip to introduce sample onto the sample absorbing material. Thepeel strip may include a peel tab at one end of the peel strip tofacilitate movement of the peel strip. The sample absorbing material maybe adapted to receive about 0.1 to about 1.0 mL of a fluid. The sampleabsorbing material may comprise a dry cellulosic material. The teststrip may include an opposed second end having a reactor detectormaterial. The test strip may include a releasing area having a mobilephase receptor for the at least one analyte. The test strip may be sizedand adapted to be enclosed within a test strip cavity. Further, the teststrip may be sized and adapted to be enclosed within a test strip cavityof a removable incubation module. Typically, the test strip is adaptedfor selecting the detection of a diagnostic test group chosen from anantibiotic analyte, toxic analyte, analyte class, a combination thereofand the like, either quantitatively, qualitatively or both.

The test zone may include at least one analyte reference line having atheoretical reflectance value. Typically, the theoretical reflectancevalue is associated with a flow parameter on the test strip. The testzone may include a first analyte reference line having a firsttheoretical reflectance value and a second analyte reference line havinga second theoretical reflectance value. The control zone may include atleast one control line having a theoretical reflectance value. Thetheoretical reflectance value may be an optical reflectance value. Acontrol zone may include a first control line having a first theoreticalreflectance value and a second control line having a second theoreticalreflectance value. The theoretical light reflectance measurement maycomprise a no-flow development theoretical value. The no-flowdevelopment value may be a reflectance value of about 85. Thereflectance value of greater than about 85 may generate a signal todeactivate the detection of the analyte.

In other examples, the flow reference area may include at least onedownstream flow reference line. The downstream flow reference line mayinclude a theoretical reflectance value after the flow reference linereceives reagent flow thereon. The flow reference area may include anintermediary flow reference line and a downstream flow reference line.The intermediary flow reference line may include a theoreticalreflectance value after the flow reference line receives reagent flowthereon. The theoretical light reflectance measurement may comprise ano-analyte pre-test development theoretical value. The test resultreference area may include at least one test line having a theoreticalreflectance value. The test result reference area may include at leastone control line having a theoretical reflectance value. The test resultreference area may include at least one test line having a theoreticalreflectance value and at least one control line having a theoreticalreflectance value. A pre-set difference between the at least one testline's theoretical reflectance value and the at least one control line'stheoretical reflectance value may activate a test result. Further, apre-set difference between the at least one test line's theoreticalreflectance value and the at least one control line's theoreticalreflectance value may trigger an error. Typically, the error withholds atest result, including generating a no-result response.

The above summary was intended to summarize certain embodiments of thepresent disclosure. Embodiments will be set forth in more detail in thefigures and description of embodiments below. It will be apparent,however, that the description of embodiments is not intended to limitthe present inventions, the scope of which should be properly determinedby the appended claims.

BRIEF DESCRIPTION OF THE DRAWINGS

Embodiments of the disclosure will be better understood by a reading ofthe Description of Embodiments along with a review of the drawings, inwhich:

FIG. 1 is a front perspective view of one embodiment of a lateral flowassay system, with an open hood illustrating cavity and base components;

FIG. 2 is a front perspective view of the lateral flow assay systemembodiment of FIG. 1, with the hood in a substantially closed position;

FIG. 3 is a front perspective view of the embodiment of FIG. 1,illustrating examples of cavity and adjustment components;

FIG. 4 is an isolated side perspective view of assay base moduleelements;

FIG. 5 is a top view of the lateral flow assay system embodiment of FIG.1 in a closed position;

FIG. 6 is a sectional view of the lateral flow assay system embodimentof FIG. 1 taken along lines 6-6, showing circuit board components;

FIG. 7 is a front perspective view of one embodiment of a lateral flowassay system and assay components;

FIGS. 7A and 7B are front perspective and rear views of lateral flowassay embodiments shown in FIG. 7;

FIG. 8 is a front perspective view of the embodiment of FIG. 7 in aclosed position;

FIG. 9 is a partial cross-section of one example of the embodimentintroduced in FIG. 7 taken along 9-9;

FIG. 10 is a front perspective view of one embodiment of a lateral flowassay system and assay components;

FIG. 11 is a front perspective view of one embodiment of a lateral flowassay system and assay components;

FIG. 12 is an isolated view of the assay illustrated in FIG. 11, showingone example of a prior analyte development before testing triggering anerror;

FIG. 13 is a front perspective view of one embodiment of a lateral flowassay system with debris on the imaging detector; and

FIG. 14 is a front perspective view of one embodiment of a lateral flowassay system having a removable assay module.

DESCRIPTION OF EMBODIMENTS

In the following description, like reference characters designate likeor corresponding parts throughout the several views. Also in thefollowing description, it is to be understood that such terms as“forward,” “rearward,” “left,” “right,” “upwardly,” “downwardly,” andthe like are words of convenience and are not to be construed aslimiting terms. It will be understood that the illustrations are for thepurpose of describing embodiments of the disclosure and are not intendedto limit the disclosure or any invention thereto.

As introduced in FIG. 1, a lateral flow assay system 1 is shown embodiedaccording to the present disclosure. Lateral flow assay system 1includes a combined reader 100 and incubator 102. Reader 100 typicallyincludes an imaging detector, such as a sensor, while incubator 102typically includes an insulated base 4. In some embodiments, theinsulated base is a removable assay module 104. Typically, reader 100first monitors an assay for one, or more, monitoring values, includingflow rate, prior analyte development and debris. In various examples, ifa proper monitoring value is detected by system 1, incubator 102incubates the assay and reader 100 generates a test result. However, ifan inconsistent monitoring value is detected, system 1 may generate ano-result response.

As shown in FIG. 1, lateral flow assay system 1 is configured to receivean assay and analyze the assay to generate a diagnostic test result.Typically, the assay is a capillary-flow test strip. However, it iswithin the sprit of this disclosure for any of the assays herein to beother lateral flow assays.

FIG. 1 shows a housing enclosing the reader 100 and incubator 102 as anintegral diagnostic unit. Other embodiments include a housing thatpartially encloses components of lateral flow assay system 1. Typically,the reader includes cavity 3 to receive the assay, and a hood 2 toenclose the assay. The housing may have an exterior and interior, andmay be opened, for instance hood 2 about member 33, to receive an assayinto cavity 3. As illustrated in FIG. 1, hood 2 may be lifted and theassay inserted into a heating cavity such as a metallic, for examplealuminum, cavity within incubator 102. Typically, cavity 3 is surroundedby insulating material, such as a plastic material, for example athermoplastic such as polyoxymethylene, known as Delrin (DELRIN is aregistered trademark of DuPont) to insulate cavity 3, and does notdeform when heated to the temperatures required for generating a testresult.

As shown in FIG. 1, hood 2 may be opened into an access position toreceive and/or remove an assay within cavity 3 of insulated base 4. Hood2 may also be configured to substantially seal cavity 3 to enclose theassay in a closed testing position. Openings 25, 26 and 27, in hood 2allow access to adjustment fasteners 11, 12 and 13 (see FIG. 8),including screws and the like, when hood 2 is in a closed position. Inother examples, adjustment fasteners may also be accessed when hood 2 ispositioned in an open access position. Typically, adjustment fastenersalign cavity 3 in relation to optics, for instance an imaging detectordescribed hereinafter, so that changes on the assay may be detected. Forexample, test strips may have multiple line developments in variousareas on the test strip, as described hereinafter and introduced in FIG.7. By allowing fine cavity adjustment with the adjustment fastenersthrough openings 25, 26 and 27, costly and cumbersome systemrecalibration may be minimized, or avoided. For instance, depending on aparticular assay, flow, test and control lines may be in a variety ofdifferent position along the assay, as explained below, which maytrigger an unexpected reflection, or transmission, value if cavity 3 isnot properly adjusted.

As introduced above, cavity 3 may be configured to receive the assay,such as a lateral flow test strip, to position and maintain the assay inan optical alignment with reader 100. In some examples, cavity 3 isshaped with an elongated channel, for instance to receive a lateral,capillary-flow test strip.

Some embodiments of reader 100 are optical analysis readers, which ofteninclude a light source and an imaging detector, for example a sensor,that is aligned such that the light from the light source shines ontothe assay and is then reflected onto the imaging sensor. An example ofoptical reader components useful in embodiments herein is described inU.S. Pat. No. 6,124,585 (Apparatus for measuring the reflectance ofstrips having non-uniform color), issued Sep. 26, 2000, and incorporatedherein by reference. Typically, the presence and, in some cases, theconcentration, of an analyte on an assay may be determined by measuring,for instance, the optical reflectance from an area of development on theassay. In some examples, percent reflectance may be used to determinethe result. In other examples, transmission may be used to detect theresult. For instance, the assay may be transparent and include a surfacehaving a transmission profile, similar to the reflectance profilediscussed below. This structure and function described in that patentmay be adapted by those of ordinary skill in the art in accordance withthe disclosure herein to obtain a functioning unit.

Reader 100 may comprise a variety of light sources, including anincandescent bulb, a fluorescent tube, a light emitting diode or thelike. In some examples, the light source may be an array of discretelight sources, for instance colored light emitting diodes chosen fromred, green, blue and a combination thereof. In yet other examples, thelight source may be an individual light source, for instance a singulardiode. Typically, the light source is configured and current driven toemit an illumination pattern suitable for reflecting onto the assay, forinstance along an elongated test strip. As shown in FIG. 1, light can bedirected to the assay, for example through aperture 5 in cavity 3, andthen reflected off the assay, back through the cavity aperture 5 anddirected to an optical detector.

In one example, an optics circuit board 31 (see FIG. 6) may have aplurality of light emitting diodes (LEDs) mounted thereon, for instancein a predetermined pattern around light-emitting aperture 5. The LEDsmay be mounted on one side of optics circuit board 31. An opticaldetector array may be mounted to the reverse side of the same opticscircuit board 31. Further, a first mirror may be positioned below thelight-emitting aperture at a pre-determined angle, for instance aboutthree hundred and fifteen degrees, to circuit board 31. A second mirrormay be positioned beneath the optical detector, for instance at an angleof about two-hundred and twenty degrees to circuit board 31, such that asubstantially 90-degree angle exists between first and second mirrors. Afocusing lens may be positioned between the first and second mirrors.Thereby, the light emitted from the LED array may illuminate an assayand then light is reflected therefrom through light-emitting aperture 5,for instance to the first mirror, from the first mirror through thefocusing lens to the second mirror, and from the second mirror onto theoptical detector. In that respect, the light striking the opticaldetector may cause the optical detector to generate a measurablevoltage. In some examples, the optical detector can output a data streamthat can be converted, for example by an on onboard central processingunit, into a series of 128 distinct one-dimensional numeric readings.The 128 readings can be taken multiple separate times and averaged.

In additional examples, a light processor may be coupled to the lightsource to actuate the light source and provide each light with theappropriate current to generate the desired emission pattern. The lightprocessor may be used to read and store data from the optical detector.The light processor may also be used to adjust the output of an array ofdiscrete light sources such that the emission pattern striking the lightdetector array has a uniform intensity. The lighting processor mayinclude data storage for the desired light-emission pattern.

Further, the light source may be an LED light source, including a red,green, blue LED device in a single package. For instance, the LED lightsource for the color sensor can also be three discrete LEDs. Similarly,a single white LED and three discrete photodiodes, with narrow bandwidthresponses at the red, green and blue wavelengths, can be used as adetector front-end.

In yet other examples, one LED is used with an optional feedback loop.The feedback loop can use a photodiode to sense light output variationfrom the single LED. If light output changes, a signal is sent so thatan appropriate adjustment can be made, for example, an increase ordecrease in current to the LED. Reflectance changes can be the result ofthe binding of a label, including color particles such as gold beads.Reflectance changes may also be a result of contaminants andinterferences in the optical path.

As seen in FIG. 2, optical window 8 may be positioned between the assayand reader 100, for instance between a test strip and a sensor.Typically, optical window 8 blocks debris from the assay fromcontaminating the imaging detector itself, or other system parts usedwith the sensor, such as lenses and mirrors. In some examples, opticalwindow 8 is clear and includes a handle so that optical window 8 isremovable from reader 100 for cleaning. In other examples, the removableoptical window may be disposable. In one example, the window materialincludes clear polyvinyl chloride (PVC) plastic. Window 8 may be mountedon a slide and inserted into reader 100 between cavity 3 and the sensor.The figures show only one removable and cleanable window to blockdebris, however, other embodiments include additional optical windowscovering to protect portions of the optics and/or incubator 102components.

Regardless of the presence of an optical window, it is possible thatdust and debris will infiltrate into reader 100, for example the opticalsensor mechanism. To provide an additional cleaning option, air inlet 6can be provided for compressed air. Air inlet 6 may be covered with atethered cap 10. In use, clear, optical window 8 is removed, andtethered cap 10 is detached. Compressed air is then blown through reader100, so that debris collected on, or near, the reader sensor is blownout through the opening previously occupied by window 8.

Some embodiments of reader 100 are programmed with multiple channels,each of which may have separate parameters associated with a relateddiagnostic test. Each channel selection parameter may include a standardcurve, a does-response curve and the like.

FIG. 3 shows cavity adjustment fastener 13 in cavity 3, and baseadjustment fasteners 11 and 12 in insulated base 4. Openings 25, 26 and27, in hood 2 allow access to adjustment fasteners 11, 12 and 13,including screws and the like, when hood 2 is in a closed position. Inother examples, adjustment fasteners may also be accessed when hood 2 ispositioned in an open access position. Typically, cavity adjustmentfastener 13 aligns cavity 3 in relation to optics, for instance animaging detector described hereinafter, so that changes on the assay maybe detected.

FIG. 4 shows one example of insulated base 4 and hood 2 in an openedaccess position. As shown, the bottom face of base 4 includes openingsfor cavity adjustment fastener 13, openings for base adjustmentfasteners 11 and 12 and light-emitting aperture 5.

FIG. 5 shows a top view of lateral flow assay 1 with hood 2 in closedtesting position. Window 8 is positioned on the side of the housing toallow the user to remove window 8 for cleaning. As introduced above, airmay be inserted through air inlet 6 to further clean debris from opticcomponents.

FIG. 6 is a bottom schematic view showing optics board 30, circuit board31 and display board 32. As shown, LEDs may be mounted on one side ofoptics circuit board 31. Further, as shown throughout the variousfigures, lateral flow assay system 1 may include user interface 7. Userinterface 7 includes an integrated circuit board 31 supporting a displayboard 32. In one example, user interface 7 allows a user to view flowdevelopment. Further, user interface 7 may allow a user to monitor asubsequent flow development after reader 100 has already detected atleast one flow development on the assay. Similarly, user interface 7 maydisplay a final test result, including a no-result response.

FIG. 7 illustrates one embodiment of hood 2 in open access position withassay 21 secured within test strip enclosure 20, which is adapted to bereceived by cavity 3. Examples of assay elements for particulardiagnostic tests having components useful for embodiments herein includethose described in U.S. Pat. No. 7,410,808, issued Aug. 12, 2008; U.S.Pat. No. 7,097,983, issued Aug. 29, 2006; U.S. Pat. No. 6,475,805,issued Nov. 5, 2002; U.S. Pat. No. 6,319,466, issued Nov. 20, 2001; U.S.Pat. No. 5,985,675, issued Nov. 16, 1999 and U.S. patent applicationSer. No. 11/883,784, filed Aug. 6, 2007, all of which are herebyincorporated herein by this reference.

Generally, lateral flow assay 21 is membrane-based test device, in whicha sample that is suspected of containing the analyte of interest isplaced at or near one end of the membrane strip. The sample is carriedto the opposite end of the membrane strip by a mobile phase thattraverses the membrane strip, for example by capillary action. Whiletraversing the membrane strip, the analyte in the test sample, if any,encounters one or more reagents. The reagents can include binders forthe analyte. Binders can be mobile and, therefore, flow with the sampleor be immobilized on the test strip as a capture agent. Depending on thetest configuration, either the analyte binder, the analyte itself, orsome other reagent in the test system, will be captured by theimmobilized capture agent and, thereby, produce a detectable signal. Thesignal can be generated by a label provided within the assay. Thedetectable signal can be measured, such as by optical reader 100.

Assay 21 may include at least one test line 40 in a test zone and atleast one control line 42 in a control zone. A theoretical reflectancevalue may be a comparison between a reflectance value at test line 40and a reflectance value at control line 42. A pre-set difference betweena theoretical reflectance value at test line 40 and a theoreticalreflectance value at control line 42 may activate lateral flow assaysystem 1, including reader 100, to generate a test result. Further, aseparate pre-set difference between a theoretical reflectance value attest line 40 and a theoretical reflectance value at control line 42 maytrigger an error. Triggering of the error may cause the microprocessorto withhold a test result, including generating a no-result response, ordeactivating reader 100 and/or incubator 102. Other embodiments includea comparison between a transmission value at test line 40 and areflectance value at control line 42.

A reflectance value on the assay that is inconsistent with thetheoretical reflectance value may indicate an inadequate flow in themobile phase on the assay. For instance, assay 21 may have a flow line44 with a corresponding theoretical light reflectance measurement. Ano-flow development value may be a reflectance value of about 85 on areflectance scale. Such an inadequate flow may trigger a detectablesignal to generate a no-result response. Additional examples includedeactivating the lateral flow assay system 1, including deactivatingreader 100 and/or incubator 102. In other examples, the flow referencearea may include both an intermediate flow reference line 46 with acorresponding theoretical reflectance value and a flow reference line44.

Similarly, a reflectance value on the assay that is inconsistent withthe theoretical reflectance value may also indicate a prior analytedevelopment on the assay. Such a prior analyte development may trigger adetectable signal to generate a no-result response. Further, if theassay is removed prior generating a test result, system 1 may generate ano-response result.

In some embodiments, assays 21 also include a coding reference componentwith a corresponding testing sequence for lateral flow assay system 1.The coding may be, for example, a color coding, a bar code, an RFID tagor the like, and may be positioned anywhere along the assay so thatdecoder sensor can decode the reference code, for example on the assay'ssurface. For instance, in some examples, the coding reference ispositioned along the distal end 45 of assay 21 as shown in FIGS. 7A and7B. Depending on the type of coding on the test strip, reader 100 mayrequire an integrated decoding sensor for example, a bar code reader, anRFID decoder or a color sensor.

Typically, the testing sequence is at least one temperature adjustmentparameter within incubator 102 and/or a channel selection of reader 100.Further, the reader test parameter may include an associated featurechosen from a standard curve, a does-response curve and the like. Otherembodiments include a variety of testing sequence parameters for theassociated diagnostic test being run on the assay.

In some examples, a color matrix, or matrices, reference coding,including a color chosen from red, blue, green and combination thereof,may be associated with a corresponding diagnostic test parameter. When acolor coding is used on assay 21, the color can be read by the readereither by a separate optical reading system or the same system thatreads the test result. That is, the assay can include a color portionthat, after enclosure within the system and test initiation, will beread by the color sensor to determine the reader channel and/or theappropriate incubator temperature. For example, a photodiode with a widedynamic range of sensitivity to red, green and blue wavelengths can beused as the detector. Red, green and blue LEDs can be used as the lightsource. Each LED can be turned on sequentially and the detector used todetermine the reflectance of each of the colors. A black surface(totally absorbent as containing no color) will produce no reflectanceof the given LEDs wavelength and, therefore, the detector will producelow output readings. A white surface will produce maximum reflectance ofall three LEDs. Various colors (depending on its content in the surfacemeasured) will produce output from the detector at varying levels.

Such color sensor components may be configured as a separate sensingcomponent within reader 1, or depending on the sensor used to read thetest strip result, a singular component that detects both development onthe test strip and color coding. In various examples, assays may becoded with a color that defines the test being run. For example, a redcolor can indicate a test strip to be used to detect beta-lactamantibiotics. Various matrices can also be delineated by the colorsystem. In the red example, after system 1 detects the red color on thetest strip, reader 100 and/or incubator 102 may be automaticallyconfigured for that specific assay 21, for example by temperatureadjustment of incubator 100 and selection of appropriate reflectancetest parameters within reader 102. Therefore, in some embodiments,system 1 may an integral diagnostic test unit that is triggered byspecific reference codings on the assay.

In other examples, the coding reference may comprise a radio frequencyidentification (RFID) tag. Such radio frequency signal transmits asignal from the tag to a decoding RFID sensor module. This signal can beused to start the analytic testing sequence, event, channel, temperatureor the like in the reader and/or incubator. Similarly, the referencecoding may be a bar code, wherein the bar code is placed on the assayand a bar code reader decodes the reference coding and the associatedtesting sequence information.

FIG. 8 shows assay 21 and assay enclosure 20 positioned within thereader, with hood 2 in a closed position. As shown, hood 2 is pivoteddown in a closed testing position, wherein a sensor in the reader is inan optical alignment with assay 21 to generate a test result or ano-result response.

In the closed testing position, incubator 102 may incubate assay 21 inan incubation environment. For instance, incubator 102 may heat and/orcool assay 21 to provide the proper incubation environment for acorresponding assay and diagnostic test. Typically, incubator 102 is incommunication to the cavity 3 and is capable of maintaining a consistenttemperature within cavity 3 either by heating or cooling at apre-defined rate. In some examples, incubator 102 includes insulatedbase 4. In other examples, incubator 102 incubates removable assaymodule 104, as described hereinafter. The incubator may be a temperatureadjustable incubator. In these examples, the temperature adjustableincubator may include a temperature control. In additional embodiments,the temperature adjustable incubator may allow for localized temperaturechanges.

Incubator 102 may include a heater. The heater may be a ceramic heater,a resister heater element and the like. Typically, cavity 3 is designedto be small so that the heater need only draw minimum current. In thatway, heating only essential areas and providing insulation around thoseareas minimizes power requirements. Use of various heating algorithmscan be useful. For example, a proportional integrated derivative (PID)can be used. In other examples, incubator 102 may compensate forlocalized temperature variations from the selected target temperature,for instance a target temperature according a corresponding testingsequence. Incubator 102 may also compensate for localized temperaturevariations with an analog, proportional control circuit. In otherexamples, incubator 102 may also compensate for localized temperaturevariations with a digital control circuit, for instance by utilizing aPID algorithm or a PID controller. Further, those of ordinary skillwould recognize that PI, PD, P or I controllers, and/or algorithms, donot preclude any of the inventions herein. For instance, temperatureadjustable incubator may include a digitally controlled potentiometer toallow the microprocessor selection of temperature. In other examples,algorithms are particularly useful when test results are affected bysmall temperature variations. Embodiments include incubator controlsystems that eliminate the need for manual adjustment by use ofembedded, digital temperature sensors and digital potentiometer thatprovides both accurate temperature reporting and a mechanism by which amicro-controller can adjust a stand-alone, analog, incubator controlcircuit.

In additional embodiments, cooling might be advantageous to reduce theincubation environment temperature, for example to stabilize theenvironment of a test medium and/or sample prior to incubation.

As shown in FIG. 9, test strip 21 may include a first end having asample absorbing material 23. Further, as introduced in FIG. 10, teststrip 21 may have a peel strip 50 to introduce sample onto sampleabsorbing material 23. Peel strip 50 may include a peel tab at one endof peel strip 50 to facilitate movement of the peel strip 50. Sampleabsorbing material 23 may be sized and configured to receive about 0.1to about 1.0 mL of a fluid. Further, sample absorbing material 23 may becomposed a dry cellulosic material. Other embodiments include othermaterials of sample absorbing material 23.

Typically, assay 21 also includes an opposed second end having a reactordetector material. Assay 21 may support a releasing area having a mobilephase receptor for the at least one analyte. Further, assay 21 may besized and adapted to be enclosed within test strip cavity 3. Similarly,assay 21 is typically sized and adapted to be enclosed, for exampleenclosed tightly, within an assay cavity 3 of a removable assay module104, as seen in FIG. 14. Typically, assay 21 is adapted for selectingthe detection of a diagnostic test group chosen from an antibioticanalyte, toxic analyte, analyte class, a combination thereof and thelike.

Reader 100 may include a sensor to monitor a test progress, for exampleon a lateral flow assay, and/or determine a test result from the lateralflow assay. The sensor is positioned relative to assay 21, so that achange on assay 21 can be detected by the sensor. Typically, the sensoris activated when the lateral flow assay is both positioned withincavity 3 and exposed to the consistent temperature within cavity 3 fromincubator 102. For example, the sensor can be activated by closing hood2 that encloses cavity 3. The sensor may include an optical detector anda microprocessor. Typically, the optical detector is aligned in anoptical path with the assay and is adapted to acquire an image detectionon the assay and is performing a continuous image detection acquisitionof the assay.

The sensor may be a single photodiode, multiple photodiodes, a linearphotodiode array, a charged couple device, a complementary metal oxidesemiconductor and a combination thereof. Therefore, at the same time asincubation and flow, or before, or after incubation and flow iscomplete, the optical sensors can monitor the assay and compare opticalreadings, such as reflectance and/or transmission readings, to determinevarious aspects including sample flow, interference with the opticalpath such as by debris in the optical path, line development and testresult. When the assay and line development falls within presetparameters, the test can continue to completion and provide a finalresult. Checking of the assay by the optical sensor prior to testcompletion can provide the user with additional confidence that the testwas processed properly.

Typically, the output may be a voltage, current or a digital outputproportional to light intensity as determined by signal conditioningcircuitry. Some examples of reader 100 include the TSL12T and TSL13Tsensors available from TAOS (Texas Advanced Optolectronic Solutions).The TSL12T and TSL13T sensors are cost-optimized, highly integratedlight-to-voltage optical sensors, each combining a photodiode and atransimpedance amplifier (feedback resistor=80 MΩ and 20 MΩrespectively) on a single monolithic integrated circuit. The photodiodeactive area is 0.5 mm×0.5 mm and the sensors respond to light in therange of 320 nm to 1050 nm. Output voltage is linear with lightintensity (irradiance) incident on the sensor over a wide dynamic range.

In some examples, the microprocessor may be in communication with theoptical detector, and in particular with the sensor. In other examples,the optical detector outputs to other logic means. Further, themicroprocessor may be adapted to signal the optical detector to performcontinuous image detection of the assay to generate the diagnostic testresult. The microprocessor may include, or have associated, memory tostore information corresponding to an imaging parameter. The memory mayinclude instructions for monitoring a pre-test analysis on the assay andfor generating a diagnostic test result on the assay.

In some embodiments having assays with coding references, as discussedherein, the optical detector may have a decoding ability to decode areference code on the assay. Thereby, the decoding sensor may therebyactive a corresponding diagnostic test in reader 100. For instance, thedecoding sensor may activate a corresponding channel in a multichannelreader 100 and/or activate a corresponding incubation temperatureprofile within incubator 102.

The decoding sensor may be a color sensor. For example, the color sensormay be a photodiode with sensitivity to wavelengths chosen from red,blue, green and a combination thereof. In such an example, a colorreading an arrangement of photodiodes, each with a specific colorfilter, is used as the decoding sensor and a white LED (which provides awide spectrum of light through the 3 bandwidths (Red, Green and Blue))is used as the light source. When the LED is turned on, the output fromeach of the photodiodes is obtained to deteiuiine the reflectance ofthat specific color. The decoding sensor may also be an RFID reader or abar code reader.

Although reference is often made herein to optical reflectance, andoptical reflectance readers, a variety of readers may be usefullyemployed including, for example, transmittance reader, fluorometers,luminometers, bar code readers, radiation detectors (such asscintillation counters), UV detectors, infrared detectors,electrochemical detectors or optical readers, such asspectrophotometers, charged coupled device (CCD) or complimentary metaloxide semiconductor (CMOS) can be used as an image sensor. An opticalreflectance reader can be programmed to analyze the test strip throughtwo-dimensional readings, rather than through the one dimensional,1×128, readings. For example, a 5×128 or 512×492 matrix of “pixels.”Such a 2-dimensional reading widens the reflectance capture area tocapture reflectance directly from the sides of the test strip.

In other examples, a transmittance reader, such as an ultravioletVisible Near-Infra red (UV-Vis-NIR) spectroscopy may provide acharacterization of the absorption, transmission, and/or reflectivity ofthe assay. For instance, such an analytical technique may measure theamount of light absorbed on the assay at a given wavelength. Those ofordinary skill in the art would appreciate that a molecule, or part of amolecule, can be excited by absorption. Typically, organic chromophoreswhich absorb strongly in the UV or visible portions of the spectrumnearly always involve multiple bonds, such as C═C, C═O or C═N. Thismolecular excitation energy may be dissipated as heat, for instancekinetic energy, by the collision of the excited molecule with anothermolecule, e.g., a solvent molecule, as the molecule returns to theground state. In other embodiments, the excitation energy may bedissipated by the emission of light via fluorescence. Regardless of theprocess, an excited molecule may possess any one of a set of discreteamounts of energy, for instance as described by the laws of quantummechanics. In examples herein, the major energy levels may be determinedprimarily by the possible spatial distributions of the electrons, and toa lesser extent by vibrational energy levels, which arise from thevarious modes of vibration of the molecule.

Therefore, in particular examples herein, absorbance measurements may bedetermined by the concentration of a solute on the assay. For instance,the progress of such a chemical reaction may be followed using aspectrophotometer in reader 100 to measure the concentration of either areactant or a product over time. In other examples, a transmissionspectroscopy may be used for solid, liquid, and gas sampling. Typically,light is passed through the assay and compared to light that has not.The resulting spectrum may depends on the path length or samplethickness, the absorption coefficient of the sample, the reflectivity ofthe sample, the angle of incidence, the polarization of the incidentradiation, and, for particulate matter, on particle size andorientation.

In some embodiments, the sensor monitors assay 21 for prior analytedevelopment before generating a test result. As shown in FIG. 12, prioranalyte development on test line 40 and control line 42 indicates anerror. A reflectance value on assay 21 that is inconsistent with thetheoretical reflectance value may indicate a prior analyte developmenton assay 21, including a pre-run assay, contaminated assay or the like.The prior analyte development may trigger a detectable signal togenerate a no-result, for instance a no-run, response and/or deactivateassay system 1. Other outputs may be indicative of the detectedcondition and are also within the scope of these inventions.

Further, the sensor may monitor flow development along assay 21 toassess whether an inadequate sample volume has been applied to assay 21,or that excess volume has been applied. For instance, prior todetermining the test result, the sensor may monitor the flow progress onassay 21 along flow line 44. In other examples, the sensor will monitorflow progress at both flow line 44 and along the assay, for instance atintermediary flow line 46. The sensor may be configured to sense whetheran adequate flow of a reagent occurred on assay 21, while assay 21 waswithin cavity 3, and/or whether one or more lines, i.e. reflectance ortransmission values, were present on assay 21 prior to contact of assay21 with the sample to be tested.

In addition, the sensor may be configured to detect whether dirt/debrisis contaminating the optical path. For instance, the sensor may monitorthe optical path for interference such as by debris. To determine that atest has run properly, or that the assay is free of dirt/debris,predetermined optical measurements, such as reflectance values ortransmission values, may be stored electronically. The preset values, orpreset parameters, can include a theoretical reflectance, ortransmission, value from an unused assay (prior to receiving reagents).Preset values may also include values may be one or more theoreticaltest lines and/or one more theoretical control lines on the assay, andmay also include a difference between the theoretical reflectance valuesfor the one or more control lines and the theoretical value for the oneor more test lines.

FIG. 13 shows one embodiment of lateral assay system 1, with debris 60over light aperture 5. In use, a reflectance value on an assay that isinconsistent with the theoretical reflectance value may indicate acontaminated optical path, such as debris 60 as shown here. Lateralassay system 1 may be adapted to generate a no-run response and/ordeactivate reader 100 and/or incubator 102 when the sensor detects suchan aberration.

In other examples, the optical detector may monitor at least onepre-test parameter after the optical detector has already acquired atleast one image detection on the assay. Similarly, optical detectorgenerates a test result from assay 21, for instance by a comparisonbetween at least two lines on the assay, for examples lines 40 and 42 ofthe test strip depicted in FIG. 7. As indicated above and in theincorporated references, optical detector may compare changes inreflectance values of two lines on the assay, for instance at least onetest line 40 and at least one control line 42.

Particular embodiments include configuring the lateral flow assay systemto allow concurrent incubation and reading of assay 21. The combinationallows sensors to be used to detect not only test results, but also tocheck parameters that might indicate whether or not flow has occurred onthe assay and that such flow caused a proper test result. That is, whilesample, including the potential analyte, or analytes, of interest, isflowing on assay 21 and binding is occurring in a mobile phase and onassay 21, the assay is being incubated. By combining reader 100 andincubator 102 into such an integral diagnostic unit 1, results can beachieved quicker than when assays, such as test strips or other testmedium, are incubated in one device and then moved to a separate devicefor reading. For instance, speed-to-result can be enhanced, for exampleto as little as less than about 60 seconds or even less than about 30seconds. Generally, such a combined system can be dynamic, sensingchanges in the assay as they occur by looking for areas of decreasedreflectance and/or transmission anywhere on the unused or not-fullydeveloped assay.

A level of protection is provided to prevent pre-run assays from beingread (for example, reader 100 will determine if line development, forinstance at flow line 44, intermediary flow line 44, test line 40 and/orcontrol line 42 occurred prior to the time when sample flow could havereached such line) and to prevent incorrect readings caused by debris,or similar interference with system optics.

Various triggers may initiate assay analysis of system 1. For example,closing of hood 2 may initiate test operation, including opticalmeasurement. Alternatively a separate switch can be used to initiatetest operation after hood 2 is closed. In either case, a first readingmay determine whether a proper assay is correctly position in thesystem. If assay 21 is detected, a reading sequence is initiated. Forexample, optical measurement, such as to detect light reflected offassay 21, can utilize values, such as average reflectance values, incertain areas of assay 21. Initially system 1 may analyze the assay todetermine if the optical path is clear of interference, such as fromdebris. Debris can be in any number of locations in the optical pathincluding on assay 21 or assay container 22. Concurrently with analyzingthe optical path for debris, or subsequent thereto, the system cananalyze the assay to determine if line development has already occurred.That is, whether a proper assay has been inserted into cavity 3. Forexample, test strips configured to develop within certain areas, such asa test line and control line, should have no development in those areasbefore the analyte and mobile phase have had adequate time to reachthem.

In some examples, lines configured to develop a change in reflectance,and/or transmission, when contacted by reagents and sample should notdevelop until flow of sample and reagents has arrived and binding hasoccurred. That flow will not have arrived at the time of an initial, forexample about three second, read. As such, if line development isdetected at the initial assay analysis, then an error message will bedelivered to the user and further readings, for example further opticalmeasurements, can be aborted. In this way, this mechanism can detect theuse of pre-run (known negative) assay or pre-marked assays. Generally,when reflectance is reduced on an unused assay, either by the presenceof line development or other darkening of the assay away from baseline,the reduction in reflectance can inform the user that something hasoccurred either on the assay or in the optical path, so that the resultshould not be accepted.

After initial optical readings are found satisfactory and appropriatereader parameters and incubator temperatures are selected, eithermanually or automatically, further optical readings, for exampleapproximately fifteen seconds after sample has been applied, can be usedto determine whether adequate flow has occurred. For example, opticalreadings can determine whether or not reagents have flowed between asample application region and a downstream line such as a test line.

The presence of label, such as colored particles, for example gold solbeads, flowing in the mobile phase, and the resulting reflectancechanges on the assay between the sample application area and a firsttest line, can inform the user that flow is occurring and return anerror message if no flow is detected. An assay lacking predictablereflectance changes might either have had no sample flow, or inadequatesample flow. Certain measurements can also indicate whether excessiveflow has occurred, as in the case where too great a volume of sample hasbeen applied to a test strip and possible reflectance change due toreagents is overwhelmed by the excessive sample volume. Reflectancechanges between the sample application area and result detection areas,such as test line and control line, can be temporary and disappear asthe mobile phase flows. If optical measurements are taken suchtemporary/non-permanent changes can be detected.

If an assay, including a test strip or other assay type, has passed thepreliminary readings, system 1 may initiate readings to generate a testresult. For example, after approximately thirty seconds test line andcontrol line analysis can begin. When there is enough differentiation,for example percent reflectance difference, between the test andcontrol, a result can be provided. Typically, negative results and moreextreme results can be provided sooner and results closer to thresholdlevels will take longer. For example, in the case of a test in which thereflectance value on the test line relates inversely to the amount ofanalyte, if the test line reflectance is reduced to a certain level thena negative result can be called. In some examples, if hood 2 is openedwhile reader 100 is reading the assay, a signal may generate a no-resultresponse.

Reader 100 and/or incubator 102 may be powered by a power source. Insome examples for on-site analysis, for instance in rugged environments,the power source may be a vehicle battery. Further, reader 100 may be incommunication with an onboard vehicle system.

As introduced in FIG. 14, lateral flow assay system 1 may includeremovable assay module 104 to be removed from system 1 and cleaned fromdebris. Typically, removable assay module104 includes a similar assaycavity as described above, to align assay 21 with optics of reader 100while in a closed testing position. In some examples, the assay is alateral flow test strip and the assay cavity within removable assaymodule 104 is sized to receive the lateral flow test strip.

As discussed above, removable assay module 104 may include a hood. Thehood may enclose the assay in a closed testing position and be opened toclean away debris in an open maintenance position when removable assaymodule104 is removed from system 1. In some examples, if the hood ofremovable assay module 104 is opened while reader 100 is reading theassay, a signal may generate a no-result response. Further, removableassay module 104 may have a bottom face having a window 108 to slide inbetween reader 100 and the assay in a manner so that at least one lightaperture 5 aligns with the assay in a closed testing position. Window108 may be removable and cleanable as discussed above, and further thebottom face may include holes to receive an adjustment fastener tosecure removable assay module 104 into an optical alignment with reader100. In other examples, bottom face 108 may include engagement lip 106to position bottom face 108 securely with reader 100.

FIG. 15 illustrates a flowchart of one testing sequence embodiment ofthe present disclosure. As shown in FIG. 15, the diagnostic test maybegin with creating an assay. For instance, creating the assay mayinclude adding a test sample to a test medium, such as a lateral flowtest strip, e.g. including any of the test strip embodiments previouslyshown or described. Typically, the test medium is configured to providea detectable test result after incubation with the test sample. Next,the assay is positioned to an optical sensor, e.g. including any of thesensor embodiments previously shown or described. The assay thenundergoes diagnostic testing concurrently with incubation as describedherein for the detection of an analyte.

In some examples as shown in FIG. 15, a clear positive or clear negativetest result, i.e. a clear positive or clear negative detection of ananalyte will end the testing sequence. However, a borderline result willtrigger a further development in the testing sequence. Typically, thefurther development includes continued performance of a diagnostic testwith concurrent incubation. In exemplary embodiments having a borderlinepreliminary result, further development is performed until a clearpositive or clear negative test result, which ends the testing sequence.

FIG. 16 illustrates a flowchart of another testing sequence embodimentof the present disclosure. As shown in FIG. 16, the diagnostic testsequence is similar to the sequence introduced in FIG. 15; however, thereader is programmed to perform only a one minute diagnostic test read.Again, a clear positive or clear negative detection of an analyte willend the testing sequence, but a borderline result will trigger a furtherdevelopment in the testing sequence. In some examples, the second oradditional diagnostic testing, e.g. any of the testing previouslydescribed herein, may also be one minute diagnostic reads. However,those of ordinary skill in the art having this disclosure will recognizeother examples include a variety of testing lengths and sequences tomeet a particular application or test result.

FIG. 17 shows another testing sequence embodiment of the presentdisclosure, where the optical sensor decodes an assay reference, e.g.any of the reference embodiments previously shown or described. Forinstance, the optical sensor may decode the assay reference to initiatea particular diagnostic test, incubation environment or the like.

FIG. 18 illustrates yet another testing sequence of the presentdisclosure.

In other embodiments, the disclosure includes a lateral flow assaysystem 1 kit. In this embodiment, the kit may comprise an incubator,e.g. any of the incubators and/or incubator components previously shownor described, and a reader, e.g. any of the readers and/or readercomponents shown or described.

In yet another embodiment of the disclosure, a method for analyteanalysis includes incubating the assay, e.g. including any of theembodiments previously shown or described, and reading the assay togenerate a test result, e.g. including any of the embodiments previouslyshown or described. In particular examples, a diagnostic test method fordetecting an analyte in a test sample includes adding a test sample to atest medium, such as a lateral flow test strip, to create an assay, thetest medium configured to provide a detectable test result afterincubation with the test sample; enclosing the test medium within ahood, the hood configured to enclose a cavity, the cavity configured toreceive the test medium and connected with a temperature control source,the temperature control source capable of maintaining a consistenttemperature; positioning a sensor, such as an optical sensor capable ofreading reflectance from the test medium, relative to the test medium sothat a change on the test medium is detectable by the sensor; andactivating the sensor, such as by closing the hood, the activationcausing the sensor to compare the test medium to a preset parameter.When the test medium is not within the preset parameter, a test resultis not provided, and wherein when the test medium is within the presetparameter, the test result is determined from the test medium, the testresult indicating whether an analyte was detected in the test sample.

In other embodiments of the methods, a preset parameter can be used todetermine either or both whether an adequate flow of reagents occurredon the test strip while the test strip was within the cavity and whetherone or more test lines are present on the test strip prior to beingcontacted by the test sample. To do so the sensor can be configured tocontinuously analyze changes on the test medium until a test resultoccurs. The test result can be determined by a comparison betweenchanges, such as reflectance changes, in a first line, for example atest line, and a second line, for example a control line, on the teststrip.

A further example of the methods include using preset parameters tocompare the test strip, prior to sample flow thereon, including prior tosample application, with the actual strip being used. For example, ablank strip, prior to reagent flow or prior to sample application, willhave a theoretical reflectance profile within a predictable range. Ifareas of reduced reflectance are detected, that did not result fromsample/reagent flow on the strip, then it is possible not only thatsomething untoward has occurred with the test strip but also it ispossible that the optical path has become contaminated and requirescleaning. Such contamination can be on the strip or within the reader.Generally, an unused test strip should have no areas of reducedreflectance. Any such areas can indicate a problem, whether fromdirt/debris, use of a test strip that was already run, or otherwise. Inany case, the test result may not be valid.

Numerous characteristics and advantages have been set forth in theforegoing description, together with details of structure and function.Many of the novel features are pointed out in the appended claims. Thedisclosure, however, is illustrative only, and changes may be made indetail, especially in matters of shape, size, and arrangement of parts,within the principle of the disclosure, to the full extent indicated bythe broad general meaning of the terms in which the general claims areexpressed. It is further noted that, as used in this application, thesingular forms “a,” “an,” and “the” include plural referents unlessexpressly and unequivocally limited to one referent.

We claim:
 1. An apparatus to generate a test result from an assay whencontacted with a sample, said apparatus comprising: a. an incubatoradapted to incubate said assay; and b. an optical detector adapted todetect a first transmission of light result on said assay and adapted todetect at least a subsequent transmission of light result on said assay,and wherein incubation of said assay and detection of said transmissionsof light on said assay generates said test result.
 2. The apparatus ofclaim 1, wherein said apparatus is adapted to perform a continuous imagedetection of said assay.
 3. The apparatus of claim 1, wherein anincubation environment comprises a heated environment.
 4. The apparatusof claim 1, wherein an incubation environment comprises a cooledenvironment.
 5. The apparatus of claim 1, wherein an incubationenvironment comprises a maintained consistent temperature environment.6. The apparatus of claim 1, wherein said optical detector detects atransmission of light passed through said assay.
 7. The apparatus ofclaim 1, wherein said optical detector monitors at least one pre-testparameter after acquiring at least one image detection on said assay. 8.The apparatus of claim 7, wherein the image detection is an opticalreflectance value.
 9. The apparatus of claim 1, wherein said assay is atest strip having at least one test line and at least one control line,and whereby a theoretical reflectance value is a comparison between areflectance value at said test line and a reflectance value at saidcontrol line.
 10. The apparatus of claim 1, wherein said incubator is atemperature adjustable incubator comprising at least one temperaturecontrol.
 11. The apparatus of claim 1, further including a housingadapted to substantially enclose said reader and said incubator.
 12. Theapparatus of claim 1, further including a user interface having adisplay board.
 13. An assembly to generate a test result from an assay,said assembly comprising: a. an optical detector adapted to image afirst transmission of light result on said assay and adapted to image aplurality of subsequent transmissions of light results on said assay;and b. an incubator adapted to incubate said assay.
 14. The assembly ofclaim 13, wherein said assembly includes a housing substantiallyenclosing said incubator and said optical detector.
 15. The assembly ofclaim 13, wherein said assembly performs continuous image detection ofsaid assay to generate said test result.
 16. An incubated apparatus togenerate a test result from an assay when contacted with a sample, areader comprising: a. an optical detector adapted to image a firsttransmission of light on said assay and adapted to image a plurality ofsubsequent transmissions of light on said assay, and wherein incubationof said assay and imaging of said transmissions of light on said assaygenerates said test result.
 17. The apparatus of claim 16, including afirst incubation environment comprising a heated environment.
 18. Theapparatus of claim 16, including a second incubation environmentcomprising a cooled environment.
 19. The apparatus of claim 16, whereinsaid incubator receives and said reader detects transmissions of lighton a test strip.
 20. The assembly of claim 16, wherein said readerperforms continuous imaging of said assay to generate said test result.